应用基因芯片筛选人高、低转移肺巨细胞癌细胞株差异表达基因的初步探讨  

Detection of differentially expressed genes between high-rate and low-rate metastatic human lung giant cell carcinoma cell strains using cDNA microarray

作　　者：梁志欣[1] 陈良安[1] 田庆[1] 王平[1] 范保星[1] 刘又宁[1] 

机构地区：[1]解放军总医院呼吸科,北京100853

出　　处：《第三军医大学学报》2006年第6期523-526,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目(30370616)~~

摘　　要：目的应用基因芯片探讨人高、低转移肺巨细胞癌细胞株95D和95C的基因表达差异,筛选肺癌转移相关基因。方法提取人高转移肺巨细胞癌细胞株95D和人低转移肺巨细胞癌细胞株95C的mRNA并反转录为cDNA,分别用Cy5-dUTP和Cy3-dUTP进行标记后与基因芯片杂交,然后用Scan Array 3000扫描仪及Im aGene3.0软件处理杂交信号得到95D和95C的表达差异基因。对部分有差异表达的基因进行RT-PCR分析鉴定。结果在被检测的人高、低转移肺巨细胞癌细胞株95D和95C中,共有466条基因出现表达差异,其中具有同一GenBank ID号的Doub le基因共108对,包括上调基因47对,下调基因61对。有表达差异的F ln29、RUVBL2、C14orf3等基因RT-PCR结果与基因芯片一致。结论多基因参与了肺癌的转移过程,基因芯片技术是筛选肺癌转移相关基因的有效方法。Objective To analyze the differences in gene expression between human lung giant cell carcinoma cell strains of high metastatic 95 D and low metastatic 95 C and to screen lung cancer metastasis-associated genes using cDNA microarray. Methods The mRNAs were extracted from human lung giant cell carcinoma cell strains of high metastatic 95D and low metastatic 95C , and reversely transcribed to the cDNAs and labeled by CyS-dUTP and Cy3-dUTP to prepare the hybridization probes. The mixed probes were hybridized to the cDNA microarray chip. The information was obtained by managing the cDNA microarray chip with Scan Array 3000 scanner and ImaGene3.0 software. A part of genes whose expressions changed in cDNA microarray analysis were further identified by RT-PCR. Results The cDNA microarray analysis showed that the expressions of 466 genes changed, among which 108 pairs of Double Gene had the same GenBank ID and some of the genes, including Fln29, RUVBL2, C14orf3 et al, were further confirmed by RT-PCR. The results of RT-PCR was coincided well with the cDNA microarray results. Conclusion Many different genes are involved in the metastasis of lung cancer, cDNA microarray technique might be a useful method in screening lung cancer metastasis-associated genes.

关 键 词：肺癌 基因芯片 基因表达 RT—PCR 

分 类 号：R394-33[医药卫生—医学遗传学] R73-354[医药卫生—基础医学]